L’utilisation de glucocorticoïdes (GC) pour le traitement de maladies inflammatoires ou d’antagonistes des androgènes pour le cancer de la prostate est limitée par l’induction d’effets secondaires, tels que l’atrophie musculaire. Comme les mécanismes sous-jacents étaient mal connus, nous avons caractérisé le rôle de ces hormones dans la régulation des fonctions musculaires. Nos résultats montrent que le récepteur aux GC des myofibres contrôle négativement la masse musculaire par des actions distinctes en présence de concentrations physiologiques et pharmacologiques de GC. De plus, notre étude a permis d’identifier de nouveaux réseaux de gènes contrôlés par les GC dans le muscle. Nous avons également démontré que les androgènes favorisent le gain de performance musculaire via l’amélioration de la force. Ainsi, cette étude a clarifié les mécanismes régulant l’homéostasie musculaire et ouvre des perspectives prometteuses pour identifier de nouvelles cibles thérapeutiques. / The use of glucocorticoids (GC) to treat inflammatory diseases or androgen antagonists for prostate cancer is limited by the occurrence of side effects such as muscle atrophy. As the underlying mechanisms were unclear, we characterized the effects of GC and androgens on muscle mass and function. Our results demonstrate that myofiber GC receptor negatively controls muscle mass by distinct actions under physiological and pharmacological levels of GC. Moreover, our data identified many genes and networks controlled by GC in myofibers. We also showed that androgens promote the gain in muscle performance during postnatal development via the improvement of specific maximal force and power. Thus, this study allowed to clarify the molecular and cellular mechanisms regulating muscle homeostasis, and paves the way to identify new therapeutic targets.

L’utilisation chronique des glucocorticoïdes conduit généralement au développement d’une résistance aux glucocorticoïdes impliquant une modulation de l’expression de gènes impliqués dans l’inflammation ou la progression tumorale comme celui codant la MMP-9. L’expression de la MMP-9 est régulée par des mécanismes épigénétiques notamment l’acétylation des histones gouvernée par la balance HDAC/HAT (histone deacetyltransferase/histone acetyltransferase) et les micro-ARNs (miRs), mécanismes également impliqués dans les phénomènes de cortico-résistance. Cependant, les altérations épigénétiques survenant suite à un traitement chronique aux glucocorticoïdes et leurs répercutions sur l’expression de la MMP-9 ne sont pas encore clairement élucidés. L’objectif de notre étude consiste donc à déterminer le rôle des mécanismes épigénétiques dans le phénomène de résistance aux glucocorticoïdes à travers leur implication dans la régulation de l’expression de la MMP-9. Nous avons montré que l’exposition chronique aux glucocorticoïdes a perturbé, à la fois, la balance HDAC/HAT (favorisant une hyperacétylation des histones) et l’expression des miRs conduisant ainsi à une surexpression de la MMP-9. De plus, nous avons montré qu’en inhibant les HDACs par le MS-275 amplifiant ainsi le phénomène d’hyperacétylation des histones, le profil d’expression des miRs a été davantage perturbé, une perturbation qui s’est répercuté sur l’expression et la sécrétion de la MMP-9. En conclusion, nous avons montré que l’acétylation des histones et la modulation de l’expression des miRs pourraient agir en harmonie pour contrôler l'expression de la MMP-9 dans un contexte de résistance aux glucocorticoïdes. / Chronic use of glucocorticoids generally leads to the development of glucocorticoid resistance which involves a modulation of the expression of genes involved in inflammation or tumor progression as that encoding the MMP-9. MMP-9 expression is regulated by epigenetic mechanisms including histone acetylation governed by the balance HDAC / HAT (histone deacetyltransferase / histone acetyltransferase) and microRNAs (miRs). These mechanisms could be involved in the phenomena of cortico-resistance. However, epigenetic alterations occurring after chronic treatment with glucocorticoids and their repercussions on MMP-9 expression are not yet clearly understood. The aim of our study is to determine the role of epigenetic mechanisms in the phenomenon of glucocorticoid resistance through their involvement in the regulation of MMP-9 expression. We demonstrated that chronic exposure to glucocorticoids disrupted both the balance HDAC / HAT (favoring a histone hyperacetylation) and expression of miRs thus leading to an overexpression of MMP-9. In addition, we have shown that inhibiting HDACs by MS-275 thereby amplifying the phenomenon of histone hyperacetylation, the expression profile of miRs was more disturbed. This disturbance has affected MMP-9 expression and secretion. In conclusion, we have shown that histone acetylation and modulation of miRs expression of harmony miRs could act to control the expression of MMP-9 in a glucocorticoid resistance context.

La grande diversité des réponses de stress observée entre individus a pour origine des facteurs génétiques en interaction avec des facteurs environnementaux. Certaines réponses peuvent être moins adaptées et accroitre la vulnérabilité de l’individu aux divers troubles et pathologies liées au stress. La CBG est une glycoprotéine plasmatique impliquée dans la biodisponibilité des glucocorticoïdes, un des principaux médiateurs de la réponse au stress. Des études génétiques ont montré que des polymorphismes du gène codant la CBG ont un impact significatif sur la réponse des glucocorticoïdes au stress. Pour comprendre les mécanismes de l’impact de la CBG sur l’action des glucocorticoïdes et les conséquences sur les réponses endocriniennes et comportementales de stress, notre équipe a développé un modèle de souris déficiente pour le gène Cbg. Ces souris présentent une réponse diminuée des glucocorticoïdes au stress, associée à un niveau élevé de comportement émotionnel de type dépressif. Cette thèse a pour but d’explorer plus en profondeur les altérations physiologiques et comportementales des souris Cbg ko. Nous avons montré que le niveau plus faible de glucocorticoïdes observé chez la souris Cbg ko provient d’une élimination plasmatique plus importante. Une étude chez la souris Cbg ko femelles a montré que les estrogènes se surimposent à la déficience en CBG pour induire des comportements de type dépressif. Nous avons également démontré que la déficience en CBG conduit a une atténuation de la sensibilité comportementale et endocrinienne au stress chronique. Enfin, nous avons observé une détérioration de la mémoire long terme de ces souris. Par ailleurs, nous montrons que dans des conditions de stress chronique associé à un régime alimentaire déséquilibré le métabolisme du glucose était altéré chez les animaux déficients en CBG. Ces résultats renforcent l’importance du rôle de la CBG influençant l’ensemble des mécanismes d’actions des glucocorticoïdes par la modulation de leurs niveaux et de leur disponibilité. / The great diversity in the response to stress observed among individuals originates from their genetic background in interaction with environmental factors. Some responses can be less adaptive and increase the vulnerability to develop stress-associated disorders. CBG is a plasma glycoprotein that regulates the bioavailability of glucocorticoids, one of the main mediators of the stress response. Genetic studies pointed out variations in the gene coding for CBG as a major factor influencing the glucocorticoid response to stress. To better understand the mechanisms involved and the consequences on endocrine and behavioral responses to stress, our team has developed a mouse model of CBG deficiency. These mice present blunted glucocorticoid response to stress associated with increased despair-like behaviors. This thesis aimed at further exploring the physiological and behavioral alterations presented by the Cbg ko mice. We showed that the lower glucocorticoid levels observed in Cbg ko mice stems from their higher clearance from plasma. A study performed on Cbg ko female mice revealed that estrogens outpass CBG deficiency in inducing despair-like behavior. Additionally, we evidenced that CBG deficiency leads to lower behavioral and endocrine sensitivity to chronic stress, and we observed impairment of hippocampal-dependent long-term memory in these mice. Finally, we found that chronic stress combined to high-fat diet leads to alteration in glucose metabolism in CBG deficient animals. These findings reinforce the important role of CBG influencing the broad range of actions of glucocorticoids by modulating their levels and availability.

As doenças inflamatórias intestinais (DII) são causadas por desequilíbrio entre as respostas imunes efetoras e reguladoras na mucosa intestinal e podem ser moduladas pelo eixo hipotálamo-hipófise-adrenal (HPA) por meio de interações neuroimunoendócrinas e secreção de cortisol. Embora os glicocorticóides (GC) sejam utilizados para tratar a DII, o cortisol produzido pelas glândulas supra-renais também está envolvido na resposta ao estresse, que pode levar a doenças inflamatórias descontroladas. Portanto, o objetivo deste trabalho é avaliar a participação do eixo HPA na modulação da resposta imune de mucosa intestinal. Para tal, camundongos C57BL/6 foram submetidos à remoção das glândulas adrenais seguida por indução de colite pela administração de água contendo 3% de dextran sulfato de sódio (DSS). Os resultados demonstraram que a ausência das adrenais levou à maior suscetibilidade à doença e mortalidade precoce, fenômeno que não foi prevenido pela reposição de GC. Os animais adrenalectomizados com colite apresentaram níveis significativamente menores de LPS, concomitantemente ao aumento de IL-6 no soro quando comparados aos camundongos não adrenalectomizados. Além disso, os animais adrenalectomizados apresentaram menor celularidade na lâmina própria (LP), menos áreas de erosão e menor escore histopatológico associado ao aumento de IFN-? e FasL, no intestino, sem produção local compensatória de corticosterona. Houve aumento na atividade das enzimas mieloperoxidase (MPO), N- acetilglicosaminidase (NAG) e eosinófilo-peroxidase (EPO) no intestino dos animais expostos ao DSS quando comparados ao grupo de camundongos controles saudáveis, independentemente da presença do eixo HPA intacto e o tratamento com GC nos animais adrenalectomizados levou à redução significativa da atividade de MPO. Também foi observado na LP dos camundongos adrenalectomizados aumento significativo na frequência de células dendríticas tolerogênicas CD11b+CD11c+CD103+, T auxiliares (CD3+CD4+), T citolíticas (CD3+CD8+) e NKT (CD3+CD49b+), além de redução significativa da população de células dendríticas pró-inflamatórias CD11b+CD11c+CD103-, leucócitos CD11b+ e linfócitos intra-epiteliais, de maneira dependente de GC. A ausência do eixo HPA intacto levou à diminuição de leucócitos totais no baço quando comparados ao grupo com colite, relacionada principalmente à redução significativa na frequência de células NKT (CD3+CD49b+), as quais foram restauradas nos camundongos tratados com GC exógenos. Durante a exposição ao DSS houve aumento de células Th2 e Th1 no baço dos camundongos não adrenalectomizados, enquanto que a remoção das adrenais levou a notável redução na população de células T CD4 produtoras de IL-4, IL-10, IFN-? ou IL-17, com aumento de células Th17 e diminuição significativa de células Th1 no baço dos camundongos adrenalectomizados e tratados com GC. De forma interessante, houve menor acúmulo de células T reguladoras juntamente à redução na intensidade média de fluorescência (MFI) de FOXP3 em células T CD4+CD25+ do baço dos camundongos adrenalectomizados expostos ao DSS, de maneira geral dependente de GC. Por fim, esta diminuição de mecanismos reguladores foi acompanhada de menor índice de proliferação e aumento de IL-10 no sobrenadante de cultura de esplenócitos de camundongos com o eixo HPA não ii funcional, indicando que a ausência de GC endógenos pode alterar significativamente a homeostase do sistema imunológico. Juntos, nossos resultados demonstram que o eixo HPA é importante na modulação da resposta imunológica durante a colite induzida experimentalmente / Inflammatory bowel diseases (IBD) are caused by imbalance between regulatory and effector immune responses in the intestinal mucosa and can be modulated by the hypothalamic-pituitary-adrenal (HPA) axis via neuroimmune endocrine interactions and secretion of cortisol. Although glucocorticoids (GC) are used to treat IBD, cortisol produced by the adrenals glands is also involved in the stress response, which can lead to uncontrolled inflammatory diseases. Therefore, the aim of this study was to evaluate the HPA axis in the modulation of the immune response of intestinal mucosa. C57BL/6 mice were subjected to removal of the adrenal glands followed by induction of colitis by administration of water containing 3% dextran sulfate sodium (DSS). The results showed that the absence of adrenals led to increased susceptibility to disease and early mortality, a phenomenon that was not prevented by GC replacement. Adrenalectomized animals exposed to DSS had significantly lower levels of LPS, concomitantly to increased IL-6 in the serum when compared to non-adrenalectomized mice. In addition, adrenalectomized animals had lower cellularity in the lamina propria (LP), less erosion areas and less histopathologic score associated with increased IFN-? and FasL in the intestine, without compensatory local production of corticosterone. There was an increase in the activity of the myeloperoxidase (MPO) enzyme, N- acetilglicosaminidase (NAG) and eosinophil-peroxidase (EPO) in the intestines of DSS-exposed animals when compared to the healthy control group of mice, regardless of the presence of intact HPA axis, while treatment with GC led to significantly reduced MPO activity. It was also observed in the LP of adrenalectomized mice significant increase in the frequency of tolerogenic dendritic cells CD11b+CD11c+CD103+, helper T (CD3+ CD4+), cytolytic T (CD3+ CD8+) and NKT (CD3+ CD49b+) besides significant reduction in the population of pro-inflammatory dendritic cells CD11c+ CD11b+ CD103-, leukocyte CD11b+ and intraepithelial lymphocytes, GC-dependent manner. The absence HPA intact carried decrease in total leukocytes in spleen when compared to the group with colitis, related mainly to significant reduction in the frequency of NKT cells (CD3+CD49b+), which were restored in the GC treated mice. During exposure to DSS there was increased Th2 and Th1 cells in the spleen of non-adrenalectomized mice, while the removal of the adrenals was associated to a marked reduction in the population of CD4 T cells producing IL-4, IL-10, IFN-? or IL-17 with increased Th17 cells and significant decrease in Th1 cells in the spleen of adrenalectomized mice treated with GC. Interestingly there was less accumulation of regulatory T cells together to a reduction in mean fluorescence intensity (MFI) of FOXP3 in CD4+CD25+ T cells in the spleen of mice exposed to DSS after adrenalectomy, most dependent on GC. Finally, the decline of regulatory mechanisms was accompanied by lower rates of proliferation and increased IL-10 in the supernatant culture of splenocytes of mice with disrupted HPA axis, indicating that the absence of endogenous GC altered significantly the homeostasis of the immune system. Together, our results demonstrate that the HPA axis is important in modulating the immune response during experimentally induced colitis

Stress reaction is usually activated by the brain, when homeostasis is or perceived to be threatened. The stress signals are transmitted from the brain by two main branches; the sympathoadrenomedullary and the hypothalamo-pituitary-adrenal (HPA) axes and employ neural, humoral and immune pathways to cope with the stressor. Because of its potency, the stress reaction has to be precisely regulated. The HPA axis is regulated by feedback loops where its end product, corticosterone in laboratory rat and mouse, inhibits its activity. The effect of corticosterone does not depend only on the concentration of corticosterone but also on local metabolism of glucocorticoids via oxo-reduction catalyzed by the enzyme 11β -hydroxysteroid dehydrogenase 1 (encoded by the Hsd11b1 gene), which intracellularly regenerates active corticosterone from inactive 11-dehydrocorticosterone, or by extra-adrenal de novo steroidogenesis of glucocorticoids. We focused on analysis of stress response in experimental animals differing in HPA axis responsivity (Fischer 344 rats (F344) vs. Lewis rats (LEW) and germ-free (GF) vs. specific pathogen free mice (SPF)) with special emphasis on regulation of stress response, glucocorticoid regeneration and influence of gut microbiota. We found that stress modulated local regeneration of...

The innate and adaptive immune processes are modulated by hormones including glucocorticoids and by microbiota. The exact mechanisms underlying the microbial and hormonal contributions to this control are not completely clear. Present study is therefore focused to crosstalk between microbiota and de novo biogenesis or local regeneration of glucocorticoids. In particular, the study analysed the effect of commensal microbiota on expression of genes encoding steroidogenic enzymes (Star, Cyp11a1, Hsd3b1, Cyp21a1, Cyp11b1) and regeneration of glucocorticoids (Hsd11b1) in adrenal glands, colon, spleen and mesenteric lymph nodes using conventional and germ-free mice. The expression of all 5 components of steroidogenesis was identified only in the adrenal gland and colon, whereas the lymphoid organs expressed predominantly Star, Cyp11a1 and Hsd3b1 indicating the ability to produce only progesterone but not corticosterone. Microbiota decreased the expression of Star in all studied tissues but the expression of other genes was insensitive to microbiota or did not respond homogenously depending on the tissue and gene. Hsd11b1 expression was upregulated by microbiota in the spleen but not in other tissues. Similarly, the in vitro treatment of immune cells isolated from mesenteric lymph nodes by microbial...

Hormones influence life of all animals. Not only they affect physiological changes in organisms, but also impact their behaviour. This work focuses at two main groups of steroid hormones: glucocorticoids and androgens. Glucocortiods are activated in response to stress. Their levels can be measured using non-invasive methods, which have a range of advantages. The main advantage is the feedback-free sample collection for enzyme immunoassay. As the measurement involves metabolites of the hormones rather than the hormones themselves, prior validation of the method is, however, necessary. This work reports on a study aiming to validate non-invasive measurement on the Madagascar Ground Gecko (Paroedura Picta). The validation was based on ACTH challenge test: Synacthen Depot was injected, which should lead to increased blood level of glucocorticoids. The validation, however, was not successful. The measurement did not discover significant increase in the levels of the metabolites of glucocorticoids. In addition, the work focuses on behavioural effects of testosterone, the primary androgen. Hormonal manipulations have been carried out on several male and female specimens. The results have discovered differences in sexual behaviour between control groups. On the other hand, the hormonal manipulations had no...

Introduction: The inflammation plays the essential role in the bone loss in juvenile idiopathic arthritis (JIA). Proinflammatory cytokines and also glucocorticoids (GCs) may activate bone resorption by osteoclasts. Simultaneously, bone formation can be attenuated, especially by inhibitors of proteins, which control the osteoblast differentiation. The aim was to verify the hypothesis that in patients with highly active JIA, reduction of bone formation via Wingless (Wnt) proteins inhibitors - Dickkopf 1 (Dkk-1) and sclerostin could be found. Except the densitometry measurements of bone and lean mass, we assessed markers of disease activity, bone metabolism and remodeling in young adult patients with JIA before and during 2 years of anti TNFα (tumour necrosis factor α) treatment, which decreases disease activity. Results: In patients with JIA before antiTNFα treatment, bone mineral density (BMD, g/cmš) was significantly reduced compared to controls. Values of BMD and body composition in JIA significantly depended on disease duration and GCs treatment. Serum concentration of sclerostin was significantly elevated in JIA compared to values in healthy controls. Values of the other monitored markers did not differ between JIA and controls. In patients with JIA, Dkk-1 correlated positively with C-reactive...

This research project explores the contribution of prenatal stress to the pathogenesis of autism as a neurobiological developmental disorder. The neurobiological impact of stress prior to the 28th week of gestation might produce structural neural changes, specifically regarding the cerebellum, the brain stem and limbic pathways, including the hippocampal area, which concept relates closely to the pathogenesis of autism. In this research project a significant focus is placed on prenatal hipothalamic-pituary-adrenal (HPA) activity due to the HPA axis’ interactivity with cortisol, digoxin and serotonin, as these biochemicals are significantly implicated in programmed foetal development, postnatal cortical behaviour, postnatal learning, as well as in functional impairment of socialization, communication and imagery associated with autism. Based upon the rationale of this research project and the conceptualisation of the topic of interest, the research problem was formulated as follows: In what unique ways does prenatal stress contribute to the pathogenesis of autism as a neurobiological developmental disorder? Sub questions included: Did the mother of the dizygotic twins experience significant stress during the period of gestation? What structural brain differences can be observed among the dizygotic twins at hand of MR-imaging? To which periods of prenatal development can these structural differences be related? How do these differences account for sensory, motor, cognitive, and affective behavioural differences among the dizygotic twins? What plasma differences can be observed among the dizygotic twins at hand of blood sampling? How does elevation of pre- and postnatal glucocorticoids relate to plasma difference among the dizygotic twins? How do these plasma differences account for sensory, motor, cognitive, and affective behavioural differences among the dizygotic twins? This research project represents quantitative research. The mode of inquiry is non-experimental at hand of a single dizygotic twin study. The following data generating strategies were employed: clinical intake interviews, administration of a diagnostic stress inventory and the 16-PF Questionnaire, MR-imaging, and the collection of blood plasma pathology results. / Dissertation (M.Ed)--University of Pretoria, 2006. / Educational Psychology / unrestricted

<p>  </p> <p>Glucocorticoids (GC) play a critical role in regulating the physiological response to stress. Disruptions to baseline levels due to stress can have negative implications on a variety of factors including growth and development, physical body conditions, metabolism, immune functions, and expression of normal behaviors, although this list is not exhaustive. When birds are unable to adapt to the stressor and return to homeostasis, the energy expenditure associated with the failed attempt at coping can lead to significant declines in the overall health, welfare, production, and performance of the bird. This can go on to impact producers and consumers as well, indicating the extensive repercussions of stress. Recently, scientists have been investigating thorough and efficient methods of quantifying stress in birds, such as measuring heterophil-to-lymphocyte ratio (HLR) or detecting glucocorticoid levels through enzyme-linked immunoassays (ELISA). However, the precise mechanism behind HLR increase during stress is unknown and ELISAs may not provide accurate results depending on when the blood is being measured. </p> <p><br></p> <p>GC are differentially released and exert their effects in a manner that is dependent on sex, age, and time. However, before investigating this, it was critical to validate the GC kits to ensure that they were measuring cortisol and corticosterone separately along with zero cross reactions with other precursors. Chapter 2 had 4 experiments carried out. The objective of experiment 1 was to validate ELISAs to ensure that they were measuring the GC accurately and separately since both cortisol and corticosterone were being measured. To do this, duck serum was pooled and charcoal-stripped to remove the presence of steroids. 3 standard curves were run to confirm that there was no cross reactivity. The objective of experiment 2 was to further validate the ELISA kits with mass spectrometry by checking for both glucocorticoids in the pooled samples. Once the validation process was complete, experiment 3 was carried out to look at the effect of ACTH stimulation on GC release. 16-week-old drakes and hens were given either intramuscular (IM) injections of cosyntropin (0.06 mg/kg) or saline as control. The cosyntropin dose was chosen according to previous studies reporting relatively high physiological responses, therefore, we wanted to replicate this. N was 10/sex/treatment. Blood was then collected at 0, 1, and 2 hours after injections and serum was analyzed by ELISAs. Lastly for experiment 4, 14-week-old developer drakes and hens at Maple Leaf Farms were assessed for a transportation stress experiment. Blood from 10 ducks/sex/time/barn were collected at 24 hours before transport to the breeder barn, immediately after a 1-hour transport, 24 hours after, and 1 week after transport. The results from experiment 1 yielded that both cortisol and corticosterone can be measured without the presence of unwanted contaminants or other products. Experiment 2 identified the greater sensitivity of mass spectrometry when reading GC levels, although the differences were linear. Experiment 3 showed that serum corticosterone levels were significantly increased at 1 hour after ACTH injections in both drakes and hens, with levels continuing to increase for the drakes. Serum cortisol levels were significantly increased at 1 hour after ACTH injections in both sexes, however, the hens had greater levels compared to the drakes. Serum cortisol levels returned to levels similar to that of saline-injected ducks at the 2-hour mark. Lastly, the transportation stress portion showed that cortisol was released at about 1/3 of corticosterone levels in both sexes. Hens showed increased levels of serum corticosterone compared to drakes at all time points except for 1 week after transport, and also had significantly increased serum cortisol levels at all time points. In conclusion, the ELISA kits were verified for future use when measuring GC as well as mass spectrometry. GC were detected in the ACTH and transportation stress experiments with hens displaying a greater sensitivity to GC release due to increased circulating levels compared to drakes. Although it was nonsignificant, there was a trend for GC to increase in response to transport. </p> <p><br></p> <p>There are sex differences in GC release and HLR for Pekin ducks and various challenges from the studies support this. With hens showing increased sensitivity to stress and drakes with more transient and gradual levels, we have consistently seen that both GC have differential roles in the stress response and not only is it critical to study both hormones, the timing of when measurements are taken are important as well to get a clear understanding of when the stress response is initiated. </p> <p><br></p> <p>Chapter 3 went further to understand the response of GC and HLR. The objective was to  investigate the release of cortisol and corticosterone in response to an ACTH dose response challenge. In Chapter 2, only one dose of cosyntropin was used and sample collection times only went to 2 hours after injections. In this study, 2 additional doses and an extra hour of sample collection were added to obtain more information. Pekin ducks were either given IM cosyntropin injections or saline for control, with an N of 10/sex/treatment. There were 3 treatment doses: High (0.06 mg/kg), medium (0.03 mg/kg), and low (0.015 mg/kg). All injections were given promptly at 0730 hours. Blood was collected at 0, 1, 2 and 3 hours after injections from the tibia veins to obtain serum for ELISAs. Blood smears were done to analyze HLR and sent to an independent lab to obtain values. The results indicated that both GC had significant sex x dose x time interactions. The low dose injection had no effect on corticosterone in hens with a slight increase for drakes at the first hour. The high dose for hens led to a spike in corticosterone levels at the first hour with a gradual decrease, and drakes had an increase that lasted for 2 hours until they returned to baseline at the last hour. The high dose in drakes stimulated cortisol release during the first 2 hours after injection with a similar effect in hens. However, hens had elevated levels compared to drakes. Finally, there was no dose response effect for HLR, although interestingly, the low dose injection elevated HLR even though there was no effect in GC. There were sex differences in the HLR response where the drakes given the high dose had levels that plateaued by the third hour, while the hens still had elevated levels. In conclusion, the ACTH dose-response test identified that ACTH has a dose-dependent effect in both GC and sex differences in their release. HLR also showed sex differences that did not depend on the dose given.</p> <p><br></p> <p>Chapter 4 observed acute exposure of GC in ducks. Pekin ducks were assigned 10/sex/treatment to receive either IM control, cortisol, or corticosterone injections. In addition, a low-dose cortisol treatment was given to represent the endogenous levels of cortisol compared to corticosterone. The control injections contained safflower oil, which was chosen as vehicle due to the low levels of genistein present. This is important as genistein is a plant estrogen and this could interact with the GC and alter the results. Blood was collected at 0, 1, 2, and 3 hours after injections for serum analysis with ELISAs, and blood smears were collected for complete blood count (CBC) differentials. Significant sex x treatment x time interactions were notable in both GC. Hens had significant increases at the first hour after injections in all treatments compared to controls, and drakes had increases at 2 hours after injections in all treatments except the low-dose cortisol. </p> <p><br></p> <p>After observing the effect of acute stress in ducks, the next step was to investigate the effects of chronic stress in chapter 5. Adult breeder Pekin ducks were randomly distributed into 3 groups: corticosterone, cortisol, or control treatments. The GC were in crystalline steroid form distributed through 2 capsules that were subcutaneously implanted on the backs of the neck. The ducks in the control group were given empty capsules. Blood smears, blood draws for serum, egg collection, body weights, and organ samples were collected over a period of 2 weeks. For the results, the corticosterone implants elevated corticosterone levels in both sexes. Interestingly, cortisol levels were elevated in both GC treatments in both sexes. Cortisol elevated HLR in drakes 1 day after implants with no effect from corticosterone. Hens had elevated HLR from both GC at all timepoints throughout the experiment. There were no significant differences in morphometrics in either sex. Corticosterone was not present in eggs, but cortisol was elevated in the albumen on day 7 and 14 of the experiment. Overall, there were sex differences in HLR where hens had greater levels in both GC treatments.</p>